Electroporation-mediated gene transfer into intact nodal meristemsin planta

Abstract

Transient expression and stable integration and expression of transgenes were observed in the tissues and offspring of certain leguminous plants after electroporation of DNA into intact nodal meristems in planta. The method described in this article thus allows the study of transgene expression in tissues differentiating from meristematic cells present in the treated buds. In addition, transgenic plants can be recovered in the offspring of electroporated individuals. Therefore, this technique allows the production of transgenic leguminous plants without the need for in vitro tissue culture, often a major hurdle with this family.

Agrobacterium-Mediated In Planta Transformation of Field Bean (Lablab purpureus L.) and Recovery of Stable Transgenic Plants Expressing the cry1AcF Gene

“…There are several reports describing legume transformation systems that require reduced or no tissue culture. Chowrira et al (1995) reported electroporation of nodal axillary buds in a range of large-seeded legumes resulting in the production of transgenic progeny. Trieu et al (2000) have described a seedling and flower infiltration method using A. tumefaciens for barrel medic that would be extremely useful in genomic studies.…”

Transient expression and stable integration of chimericGusgene in watermelon following electroporation

“…Batches of 25 embryos were suspended in electroporation buffer prepared in 1ml sterile MS medium, pH 3.7 containing 200 µg ml -1 of plasmid DNA, with DNA protectant spermine at 2 mM concentration (Akella and Lurquin, 1993;Chowrira et al, 1995). Negative controls were performed without DNA but only MS-salts and spermine (2 mM).The DNA treated embryos, suspended in the electroporation buffer taken in the circular electrode were subjected to 5 and 10 pulses of 400V/per cm pulse strength.…”

“…The plasmid DNA (200 µg ml -1 ), pBI121 or p35SGUSINT was complexed with spermine (2mM) or Lipofectin™ for 20 min in sterile MS medium (Chowrira et al, 1995). 5 ul of the DNA solution were injected from the top through the apical dome to a depth of about 1 mm using a Hamilton syringe.…”

Asiatic Beans

“…Electroporation-mediated gene transfer system described by Chowira et al (1995Chowira et al ( , 1996 uses intact nodal meristems and relies on the plant to develop its reproductive structures from the treated apices in a more or less normal fashion. Transformation of intact embryonic explants by electroporation (Dillen et al, 1995) obtained high frequency of transformation with wider applicability to diverse legume species.…”